Design of carrier peptide-oligonucleotide conjugates with rapid membrane translocation and nuclear localization properties.
Peptides containing a hydrophobic motif associated with a nuclear localization signal separated by various linkers were synthesized in solid phase. The hydrophobic sequence corresponds either to a signal peptide sequence or to a fragment of the fusion peptide of GP41 while the hydrophilic sequence is that of a nuclear localization signal. The C-termini of these peptides bear a cysteamide group that was linked to a fluorescent probe. This allowed the cellular localization of the probe to be determined as a function of the peptide sequences. The labeled peptides were then incubated with fibroblasts. Using N-biotinylated derivatives we confirmed by indirect immunofluorescence that the observed localizations corresponds to those of the peptides. The presence of a linker appears to play a role in the cellular localization. One of these peptides was successfully used to target fluorescent oligodeoxynucleotides into living cells demonstrating improved cell delivery of peptide-oligodeoxynucleotide conjugates.
Chaloin
L
L
Centre de Recherche de Biochimie Macromoléculaire, ERS 155 1919, Montpellier, France.
Vidal
P
P
Lory
P
P
Méry
J
J
Lautredou
N
N
Divita
G
G
Heitz
F
F
D000595:Amino Acid Sequence; D001692:Biological Transport / Q000502:physiology; D019921:Biotinylation; D015220:Calcium Channels / Q000235:genetics; D004705:Endocytosis / Q000502:physiology; D005347:Fibroblasts; D005455:Fluorescent Antibody Technique; D005456:Fluorescent Dyes / Q000138:chemical synthesis / Q000378:metabolism; D015700:HIV Envelope Protein gp41 / Q000737:chemistry; D006801:Humans; D008856:Microscopy, Fluorescence; D008969:Molecular Sequence Data; D019913:Nuclear Localization Signals / Q000502:physiology*; D009838:Oligodeoxyribonucleotides / Q000378:metabolism*; D016376:Oligonucleotides, Antisense / Q000378:metabolism; D018408:Patch-Clamp Techniques; D021382:Protein Sorting Signals / Q000138:chemical synthesis* / Q000378:metabolism
D015220:Calcium Channels; D005456:Fluorescent Dyes; D015700:HIV Envelope Protein gp41; D019913:Nuclear Localization Signals; D009838:Oligodeoxyribonucleotides; D016376:Oligonucleotides, Antisense; D021382:Protein Sorting Signals